The NNT Knockout Raji Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the NNT gene in the Raji human B lymphocyte cell line. This loss-of-function model enables the study of mitochondrial nicotinamide nucleotide transhydrogenase (NNT) function without clonal selection, providing a heterogeneous knockout pool that reflects population-level gene disruption. NNT encodes a mitochondrial inner membrane enzyme critical for NADPH homeostasis, and its inactivation allows researchers to dissect redox-dependent processes in a B-lymphocyte context. The polyclonal format offers a practical tool for functional genomics and oxidative stress research, avoiding the limitations of single-cell-derived clones.
The Raji host cell line is a well-characterized model derived from a Burkitt’s lymphoma patient, representing a B lymphocyte at a mature stage of differentiation. These cells are widely employed in immunology and cancer biology for studies of antibody production, antigen presentation, tumorigenesis, and immune surveillance. As a suspension cell line with robust growth characteristics, Raji cells are amenable to a range of biochemical and genetic manipulations. Their origin from a hematological malignancy also makes them particularly relevant for investigating pathways that link metabolism, redox balance, and cell survival in lymphoma and leukemia.
NNT encodes a mitochondrial inner membrane transhydrogenase that uses the proton gradient to generate NADPH from NADH, serving as a primary source of NADPH for the glutathione and thioredoxin systems. Its expression is regulated by PGC-1?? and NRF2, which coordinate mitochondrial biogenesis and antioxidant responses. The NADPH produced supports glutathione reductase (GSR)-mediated GSH regeneration, peroxiredoxin 3 (PRDX3) function, and thioredoxin reductase 2 (TXNRD2) activity, thereby maintaining mitochondrial redox balance and protection against ROS.
In the Raji B-lymphocyte context, NNT knockout is expected to impair mitochondrial NADPH generation, rendering cells more susceptible to oxidative challenges and potentially affecting processes such as proliferation, immunoglobulin production, and response to apoptotic stimuli. The model is valuable for dissecting how mitochondrial redox status influences B-cell biology and for investigating the contribution of NNT to lymphomagenesis. Moreover, because loss-of-function mutations in NNT are linked to familial glucocorticoid deficiency, these polyclonal knockout cells serve as a human cellular model for this endocrine disorder, allowing exploration of NNT’s role in adrenal physiology and metabolic stress.
These polyclonal knockout cells support diverse experimental applications, including studies of mitochondrial redox regulation in B lymphocytes, NNT’s role in survival and proliferation under oxidative stress, and screening of NNT-targeted compounds. Standard assays include western blotting, RT-qPCR, NADPH/NADP+ ratio measurements, MitoSOX ROS detection, GSH/GSSG quantification, and viability testing following H?O? challenge. Additional methods encompass Annexin V/PI apoptosis assays and metabolic flux analysis. For further details, please contact Ascent Research.
